Stabilized anti-cancer cold atmospheric plasma (cap)-stimulated media and methods for preparing and using the same

Abstract

This disclosure relates to stabilized anti-cancer cold atmospheric plasma (CAP)-stimulated media, to methods for preparing such media, and to methods of treatment using such media.

1 . A stabilized cold atmospheric plasma-stimulated anti-cancer medium, 
     
     
         2 . The medium of  claim 1 , wherein the medium comprises phosphate buffered saline (PBS), Dulbecco's Modified Eagle Medium (DMEM), or a combination thereof. 
     
     
         3 . The medium according to  claim 1 , wherein the medium comprises phosphate buffered saline (PBS). 
     
     
         4 . The medium according to  claim 1 , wherein the medium comprises Dulbecco's Modified Eagle Medium (DMEM). 
     
     
         5 . The medium according to  claim 1 , wherein the medium is stable for a period of up to 7 days, up to 6 days, up to 5 days, up to 4 days or up to 3 days. 
     
     
         6 . The medium according to  claim 1 , wherein the medium is stable for between about 1 and about 7 days, about 1 and about 6 days, about 1 and about 5 days, about 1 and about 4 days or about 1 and about 3 days. 
     
     
         7 . The medium according to  claim 1 , wherein the medium is stable at a temperature of between about −25° C. and about 25° C., between about − 25 ° C. and about 22° C., between about 0° C. and about 22° C., between about 0° C. and about 8° C. or between about 2° C. and about 8° C. 
     
     
         8 . The medium according to  claim 1 , wherein the medium is free of cysteine, methionine, or a combination thereof. 
     
     
         9 . The medium according to  claim 1 , wherein the medium is free of phenylalanine. 
     
     
         10 . The medium according to  claim 1 , wherein the medium is free of phenol red. 
     
     
         11 . The medium according to  claim 1 . wherein the medium comprises 3-nitro-L-tyrosine. 
     
     
         12 . The medium according to claim herein the 3-nitro-L-tyrosine is present in the plasma-stimulated media at a concentration of up to 9 mM. 
     
     
         13 . The medium according to  claim 11 , wherein the 3-nitro-L-tyrosine is present in the plasma-stimulated media at a concentration of between about 1 and about 5 mM. 
     
     
         14 . The medium according to  claim 11 , wherein the 3-nitro-L-tyrosine is present in the plasma-stimulated media at a concentration of about 1 mM, about 2 mM. about 3 MM. or about 4 mM. 
     
     
         15 . A method of stabilizing and/or enhancing the anti-cancer activity of cold atmospheric plasma-stimulated media, the method comprising:
 (i) reducing the amount of cysteine, methionine, or a combination thereof, in the media;   (ii) reducing the amount of phenylalanine in the media;   (iii) reducing the amount of phenol red in the media;   (iv) adding 3-nitro-tyrosine to the media; or   (v) any combination of (i)-(iv).   
     
     
         16 . A method of treating a target tissue comprising administering to a patient in need of such treatment a medium according to  claim 1 . 
     
     
         17 . The method according to  claim 16 , wherein the tissue is cancerous tissue. 
     
     
         18 . The method according to  claim 16 , wherein the tissue comprises lung tissue, bladder tissue, brain tissue, skin tissue, or any combination thereof. 
     
     
         19 . A method of enhancing the anti-cancer activity of cold atmospheric plasma—
 stimulated media, the method comprising; 
 (i) increasing the diameter of the well in a multi-plate well, 
 (ii) decreasing the gap between the plasma tube and the surface of the media during the cold atmospheric plasma treatment; or 
 (iii) any combination of (i) and (ii). 
 
     
     
         20 . The method according to  claim 19 , wherein the diameter of the well is increased by about 42% or about 57%. 
     
     
         21 . The method according to  claim 19 , wherein the gap is decreased by about 25%, about 20%, about 17%, or about 14%.